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Effect of hydrogen peroxide and dithiothreitol on contractile function of single skeletal muscle fibres from the mouse

Francisco H. Andrade *¹, Michael B. Reid ², David G. Allen ³ and Håkan Westerblad *

1. We used intact single fibres from a mouse foot muscle to study the role of oxidation-reduction in the modulation of contractile function.

2. The oxidant hydrogen peroxide (H₂O₂, 100-300 µM) for brief periods did not change myoplasmic Ca²⁺ concentrations ([Ca²⁺]_i) during submaximal tetani. However, force increased by 27 % during the same contractions.

3. The effects of H₂O₂ were time dependent. Prolonged exposures resulted in increased resting and tetanic [Ca²⁺]_i, while force was significantly diminished. The force decline was mainly due to reduced myofibrillar Ca²⁺ sensitivity. There was also evidence of altered sarcoplasmic reticulum (SR) function: passive Ca²⁺ leak was increased and Ca²⁺ uptake was decreased.

4. The reductant dithiothreitol (DTT, 0·5-1 mM) did not change tetanic [Ca²⁺]_i, but decreased force by over 40 %. This was completely reversed by subsequent incubations with H₂O₂. The force decline induced by prolonged exposure to H₂O₂ was reversed by subsequent exposure to DTT.

5. These results show that the elements of the contractile machinery are differentially responsive to changes in the oxidation-reduction balance of the muscle fibres. Myofibrillar Ca²⁺ sensitivity appears to be especially susceptible, while the SR functions (Ca²⁺ leak and uptake) are less so.

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